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A rapid assay for on-site monitoring of infliximab trough levels: a feasibility study

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Abstract

Monitoring levels of biologicals against tumor necrosis factor (TNF) has been suggested to improve therapeutic outcomes in inflammatory bowel diseases (IBDs). This pilot study describes a rapid lateral flow (LF)-based assay for on-site monitoring of serum trough levels of humanized monoclonal antibody infliximab (IFX). The applied chromatographic method utilizes sequential flows of diluted serum, wash buffer, and an immunoglobulin generic label on LF strips with a Test line comprised of TNF-α. The successive flows permitted enrichment of IFX at the Test line before the label was applied. The label, luminescent upconverting phosphor (UCP) particles coated with protein-A, emits a 550-nm visible light upon excitation with 980-nm infrared light. IFX concentrations were determined through measurement of UCP fluorescence at the Test line. The assay was optimized to detect IFX levels as low as 0.17 μg/mL in serum. For patients with IBD, this limit is appropriate to detect levels associated with loss of response (0.5 μg IFX/mL). The assay was evaluated with clinical samples from patients with Crohn’s disease and correlated well within the physiologically relevant range from 0.17 to 10 μg/mL with an IFX-specific ELISA. Performance of the assay was further successfully validated with samples from blood donors, IFX negative IBD patients, and rheumatoid arthritis patients that had developed anti-IFX antibodies. Because of its generic nature, the assay is suited for detecting most therapeutic anti-TNF-α monoclonal antibodies.

A rapid lateral flow based assay to determine trough levels of infliximab and other anti‐TNF‐α antibodies. The rapid format showed excellent and quantitative correlation with ELISA. Accurate quantitation was achieved utilizing the up‐converting phosphor reporter technology and a portable lightweight ESEQuant LFR reader adapted with an infrared LED

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Abbreviations

ADA:

Adalimumab

ATI:

Antibodies to infliximab

CF:

Consecutive flow

HSLF:

High salt lateral flow

IBD:

Infectious bowl disease

IFX:

Infliximab

Ig:

Immunoglobulin

LF:

Lateral flow

NHS:

Normal human serum

POC:

Point-of-care

RA:

Rheumatoid arthritis

RFU:

Relative fluorescent unit

TNF-α:

Tumor necrosis factor alpha

UCP:

Upconverting phosphor

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Acknowledgments

This study was financially supported by a research grant from Centocor BV, Leiden, the Netherlands and U.S. National Institute of Health grant UO1DE017855. We thank J.M. van der Zon and A.E. van de Meulen-de Jong (LUMC, Department of Gasteroenterolgy and Hepatology) for their help with the patient materials. We also thank D. van der Kleij (Sanquin, Diagnostic Services Division) for the ELISA data.

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Authors and Affiliations

  1. Department of Molecular Cell Biology, Leiden University Medical Center, Einthovenweg 20 Bldg 2, PO Box 9600, 2300 RC, Leiden, The Netherlands

    Paul L. A. M. Corstjens, Karien C. Wiesmeijer, Claudia J. de Dood & Hans J. Tanke

  2. Department of Gastroenterology, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX, Utrecht, The Netherlands

    Herma H. Fidder

  3. Sanquin Research CLB and Landsteiner Laboratory, Academic Medical Center, Plesmanlaan 125, 1066 CX, Amsterdam, The Netherlands

    Theo Rispens & Gert-Jan Wolbink

  4. Center for Inflammatory Bowel Diseases, University of California, 10945 Le Conte Ave, Los Angeles, CA, 90095, USA

    Daniel W. Hommes

  5. Department of Gastroenterology and Hepatology, Leiden University Medical Center, Postbus 9600, 2300 RC, Leiden, The Netherlands

    Herma H. Fidder & Daniel W. Hommes

Authors
  1. Paul L. A. M. Corstjens
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  2. Herma H. Fidder
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  3. Karien C. Wiesmeijer
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  4. Claudia J. de Dood
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  5. Theo Rispens
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  6. Gert-Jan Wolbink
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  7. Daniel W. Hommes
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  8. Hans J. Tanke
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Corresponding author

Correspondence to Paul L. A. M. Corstjens.

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Paul L.A.M. Corstjens and Herma H. Fidder contributed equally to this work.

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Corstjens, P.L.A.M., Fidder, H.H., Wiesmeijer, K.C. et al. A rapid assay for on-site monitoring of infliximab trough levels: a feasibility study. Anal Bioanal Chem 405, 7367–7375 (2013). https://doi.org/10.1007/s00216-013-7154-0

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  • DOI: https://doi.org/10.1007/s00216-013-7154-0

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